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. 2000 Feb 29;97(6):2491–2496. doi: 10.1073/pnas.050586597

Figure 4.

Figure 4

Reversibility of Ace1 inhibition by DEA/NO in vivo and in vitro. (A) 217CUP late log phase cells were incubated with (open bars) or without (closed bars) DEA/NO (1 mM) for 10 min at 30°C with shaking (275 rpm) in the absence (left pair of columns) or presence (middle pair of columns) of 10 mM 2-ME. The pair of columns on the right represents experiments in which exposure to 10 mM 2-ME (10 min) occurred after DEA/NO exposure. The cells were induced (CUP1-lacZ) for 60 min and were assayed for β-galactosidase activity as described in Experimental Procedures. (B) Cell extracts of 217ACE were incubated with (lanes 2, 4, and 6) or without (lanes 1, 3, and 5) DEA/NO (4 mM). To the mixtures were added 10 mM 2-ME either during (lanes 3 and 4) or after (lanes 5 and 6) exposure to DEA/NO, and they incubated for 10 min. The EMSA was performed as described in Experimental Procedures. Representative experiments are shown.